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Quantitative 3d Analysis Of Intra-Nuclear Organization In The Tissue Context

Published online by Cambridge University Press:  02 July 2020

S.J. Lockett
Affiliation:
Life Sciences Division, Ernest Orlando Lawrence Berkeley National Laboratory, CA94720
D.W. Knowles
Affiliation:
Life Sciences Division, Ernest Orlando Lawrence Berkeley National Laboratory, CA94720
D. Pinkel
Affiliation:
UCSF Cancer Center, 2340 Sutter St, San Francisco, CA94143
C. Ortiz de Solórzano
Affiliation:
Life Sciences Division, Ernest Orlando Lawrence Berkeley National Laboratory, CA94720
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Extract

Confocal microscopy is revealing associations between the internal organization of the nucleus and tissue architecture and function. Such associations may exist which are too subtle or complex for visual observation or quantitative analysis may be required, for example as input data to mathematical modeling of cellular processes. In these circumstances, it is necessary to perform the analysis using computer algorithms. We have developed 3D image analysis (IA) algorithms for segmenting nuclei from within intact tissue specimens, measuring the structure of the nuclei and for segmenting specifically- labeled punctate entities within nuclei. In this study we developed algorithms for measuring the spatial organization of the two copies of a specific DNA locus (labeled using fluorescence in situ hybridization (FISH) ) inside diploid nuclei and with respect to the nuclear organization of the tissue.

For each segmented nucleus, its center of mass (CoM) was determined, which informed us about its position in the tissue.

Type
Cytochemistry
Copyright
Copyright © Microscopy Society of America

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